Functional and hyperfunctional siRNA directed against Bcl-2

Abstract

Efficient sequence specific gene silencing is possible through the use of siRNA technology. By selecting particular siRNAs by rationale design, one can maximize the generation of an effective gene silencing reagent, as well as methods for silencing genes.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of the filing dates of U.S. patent application Ser. No. 10 / 714,333, filed Nov. 14, 2003 entitled “Functional and Hyperfunctional siRNA,” U.S. Provisional Application Ser. No. 60 / 502,050, filed Sep. 10, 2003, entitled “Methods for Selecting siRNA,” and U.S. Provisional Application Ser. No. 60 / 426,137, filed Nov. 14, 2002, entitled “Combinatorial Pooling Approach for siRNA Induced Gene Silencing and Methods for Selecting siRNA,” the entire disclosures of which are hereby incorporated by reference into the present disclosure. REFERENCE TO TABLES SUBMITTED IN ELECTRONIC FORM [0002] Applicants hereby incorporate by reference the material submitted previously, in duplicate on the compact disks labeled DOCKET 13499, PATENT APPLICATION, DISK 1 of 1, COPY 1 of 2; and DOCKET 13499, PATENT APPLICATION, DISK 1 of 1, COPY 2 of 2; which copies are identical, in files entitled Table—12.txt, date of creation Jun. 26,...

AI Technical Summary

Benefits of technology

[0012] The present invention is directed to increasing the efficiency of RNAi, particularly in mammalian systems. Accordingly, the present invention provides kits, siRNAs and methods for increasing siRNA efficacy.

[0068] The ability to use the above algorithms, which are not sequence or species specific, allows for the cost-effective selection of optimized siRNAs for specific target sequences. Accordingly, there will be both greater efficiency and reliability in the use of siRNA technologies.

Problems solved by technology

However, these attempts to induce RNAi met with limited success, due in part to the induction of the interferon response, which results in a general, as opposed to a target-specific, inhibition of protein synthesis.

Thus, long dsRNA is not a viable option for RNAi in mammalian systems.

One of the most contentious issues in RNAi is the question of the necessity of siRNA design, i.e., considering the sequence of the siRNA used.

Unfortunately, due to the interferon response, this same approach is unavailable for mammalian systems.

While this effect can be circumvented by bypassing the Dicer cleavage step and directly introducing siRNA, this tactic carries with it the risk that the chosen siRNA sequence may be non-functional or semi-functional.

Unfortunately, none of the reported methods have provided a satisfactory scheme for reliably selecting siRNA with acceptable levels of functionality.

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